GSM6729711: AS-01; Homo sapiens; RNA-Seq
Whole blood was diluted with 1× PBS (1:1), layered on top of Ficoll-Paque Plus (Cytiva, USA), and centrifuged at 400 g for 30 min. The PBMC layer was isolated and washed twice with Dulbecco's phosphate buffered saline (DPBS) at 300 g for 10 min, followed by the removal of erythrocytes with red blood cell lysate on ice. After centrifugation, the cells were resuspended in RPMI1640 containing 10% fetal bovine serum (FBS). Cell viability was verified using trypan blue and approached 90% for each sample. Cells with high viability were subjected to library preparation. RNA libraries were generated by 10xGenomics Single Cell 3' V3.1 Library Gel Bead Kit and prepared for sequencing using standard Illumina protocols